The recent success of small-molecule ATP-competitive kinase inhibitors such as Gleevec, Iressa, and Tarceva has attracted significant attention and protein kinases now represent one of the most important groups of targets for drug discovery and development. Because of their genetic and physiological similarity to humans, zebrafish show promise as an efficient animal model for drug screening. Although all members of the mammalian protein kinases have not yet been identified in zebrafish, in situ hybridization experiments showed that Src family of tyrosine kinases src, fyn and yes are ubiquitously expressed in zebrafish embryos. In this research, we will develop a high throughput in vivo zebrafish ELISA format for identifying Src family kinase inhibitors. The zebrafish assay permits evaluation of the therapeutic potential of a compound in a complex physiological environment using throughput similar to cell-based assays. Advantages of the zebrafish assay include: short assay time, low cost, ease of embryo maintenance during drug delivery, compatibility of assay procedures with standard microplate formats, single drug dosing for each embryo, and the ability to test large numbers of embryos for statistically significant results. PUBLIC HEALTH RELEVANCE: In this research, we will develop a high throughput in vivo zebrafish ELISA format for identifying Src family kinase inhibitors. [unreadable] [unreadable] [unreadable]